Ovarian carcinoma SKOV3 cell lines were obtained from the Shanghai Institute of Cell Library (Shanghai, China). The small interfering RNA (siRNA) was designed and constructed as a vector (plasmid without vector and bacterial strain containing this plasmid were all produced and identified by Nanjing Moji Biological Technology Development Co. Ltd., Nanjing, China). The main reagents used were: CCK-8, IL-8, RPMI-1640 basal culture medium, double-antibody and fetal bovine serum (FBS) (both from Gibco, Grand Island, NY, USA), acrylamide, methylene acrylamide, Tris-base, ammonium peroxydisulfate, TEMED, glycine and SDS (all from Biosharp, Hefei, China). The PVDF membrane, and chemiluminescent liquid were purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA).
Glycine
Manufactured by Biosharp
Sourced in China
Glycine is a versatile chemical compound commonly used in various laboratory applications. It is the simplest amino acid, consisting of a single hydrogen atom attached to the central carbon atom. Glycine serves as a building block for proteins and is involved in various biochemical processes. Its core function is to provide a basic structural unit for the synthesis of proteins and peptides, which are essential in biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Tris base, by Biosharp (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Sodium dodecyl sulfate (sds), by Biosharp (2 mentions)
Double antibody, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Thermo Fisher Scientific (1 mentions)
Cck 8, by Thermo Fisher Scientific (1 mentions)
Acrylamide, by Biosharp (1 mentions)
Tetramethylethylenediamine, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Biosharp (1 mentions)
Protein lysis buffer, by Thermo Fisher Scientific (1 mentions)
Molecular weight marker, by Thermo Fisher Scientific (1 mentions)
Skimmed milk powder, by Biosharp (1 mentions)
Tween 20, by Biosharp (1 mentions)
Trypsogen, by Merck Group (1 mentions)
Tris base, by Merck Group (1 mentions)
Acrylamide, by Merck Group (1 mentions)
Glycogen assay kit, by Nanjing Jiancheng (1 mentions)
Bromophenol blue, by Thermo Fisher Scientific (1 mentions)
Urea, by Bio-Rad (1 mentions)
Agarose, by Biosharp (1 mentions)
4 protocols using glycine
1
SKOV3 Cell Line Knockdown Protocol
2
Comprehensive Protein Analysis Workflow
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Each of the chemical reagents and the HP‐20 micro resin were of analytical grade and purchased from Sinopharm (Shanghai, China). Glycine, bovine insulin, sodium dodecyl sulfate, sodium salt (SDS), glycerine, tween‐20, ammonium persulfate and skimmed milk powder and dimethyl sulfoxide were purchased from BioSharp (Hefei, Anhui, China). Trypsogen, acrylamide, Tris‐base and tetramethyl ethylene diamine were purchased from Sigma (St. Louis, MO, USA). Glucose and a glycogen assay kit were purchased from Jiancheng (Nanjing, Jiangsu, China). Protein lysis buffer and the molecular weight marker were obtained from Thermo Fisher Scientific (Waltham, MA, USA). All of the antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
3
Two-dimensional Proteomic Analysis of Cell Lines
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Dulbecco’s Modified Eagle’s Medium (DMEM) was purchased from Gibco (Waltham, MA, USA), and fetal bovine serum (FBS) was also purchased from Gibco. Urea, CHAPS zwitterionic detergent (CHAPS), dithiothreitol (DTT), immobilized pH gradient (IPG) buffer, Tris-HCl, and IPG strips (17 cm, pH 4–7, linear) were purchased from Bio-Rad (Hercules, CA, USA). Collagenase (Type I), protease inhibitory cocktail, and thioUrea were purchased from Sigma-Aldrich (St Louis, MO, USA). Sodium dodecyl sulfate (SDS), glycerol, Tris base, agarose, glycine, and Coomassie R-250 were purchased from Biosharp (Hefei, China). Iodoacetamide was purchased from TCI (Portland, OR, USA). Bromophenol blue was purchased from Fisher Scientific (Pittsburgh, PA, USA). TRIzol reagent and reverse transcription kit were purchased from Takara (Shiga, Japan). Fast SYBR Green Master Mix was purchased from Toyobo (Osaka, Japan).
4
EDU Staining for Cell Proliferation
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EdU (5-ethynyl-2′-deoxyuridine) staining enables quick and effective cell proliferation assays to accurately measure the proportion of cells in the S phase. Therefore, using EdU staining in accordance with the manufacturer’s instructions, the proliferation of pGCs was determined. After treatment with 0 mmol/L, 20 mmol/L, 40 mmol/L, and 60 mmol/L 2,5-HD for 24 h, cells were cultured in phenol red-free DMEM/F12 with 50 μM EdU (RiboBio, Guangzhou, China) staining for 2 h [23 (link),24 (link)]. Cells were then rinsed two times with 0.02 M PBS (pH = 7.2) at 5-min intervals before being immobilized for 30 min with 4% paraformaldehyde (Biosharp, Hefei, China). Following that, cells were treated for 5 min with glycine (Biosharp, Hefei, China) (2 mg/mL) and rinsed for 5 min with PBS [23 (link),24 (link)]. After 30 min of DAPI (0.5 μg/mL) (Biosharp, Hefei, China) staining [23 (link)], three rounds of PBS washing were conducted at 5-min intervals. Lastly, a Leica DM4000 BLED microscope (Leica Microsystems DM2500, GER) was used to observe the fluorescence in cells.
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